Hamill Kevin J, Hopkinson Susan B, DeBiase Philip, Jones Jonathan C R
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611, USA.
Mol Biol Cell. 2009 Jun;20(12):2954-62. doi: 10.1091/mbc.e09-01-0051. Epub 2009 Apr 29.
alpha6beta4 integrin, a component of hemidesmosomes, also plays a role in keratinocyte migration via signaling through Rac1 to the actin-severing protein cofilin. Here, we tested the hypothesis that the beta4 integrin-associated plakin protein, bullous pemphigoid antigen 1e (BPAG1e) functions as a scaffold for Rac1/cofilin signal transduction. We generated keratinocyte lines exhibiting a stable knockdown in BPAG1e expression. Knockdown of BPAG1e does not affect expression levels of other hemidesmosomal proteins, nor the amount of beta4 integrin expressed at the cell surface. However, the amount of Rac1 associating with beta4 integrin and the activity of both Rac1 and cofilin are significantly lower in BPAG1e-deficient cells compared with wild-type keratinocytes. In addition, keratinocytes deficient in BPAG1e exhibit loss of front-to-rear polarity and display aberrant motility. These defects are rescued by inducing expression of constitutively active Rac1 or active cofilin. These data indicate that the BPAG1e is required for efficient regulation of keratinocyte polarity and migration by determining the activation of Rac1.
α6β4整合素是半桥粒的一个组成部分,它还通过Rac1向肌动蛋白切割蛋白丝切蛋白发出信号,在角质形成细胞迁移中发挥作用。在此,我们检验了以下假设:β4整合素相关的斑珠蛋白,大疱性类天疱疮抗原1e(BPAG1e)作为Rac1/丝切蛋白信号转导的支架发挥作用。我们构建了BPAG1e表达稳定敲低的角质形成细胞系。敲低BPAG1e不影响其他半桥粒蛋白的表达水平,也不影响细胞表面表达的β4整合素的量。然而,与野生型角质形成细胞相比,BPAG1e缺陷细胞中与β4整合素结合的Rac1的量以及Rac1和丝切蛋白的活性均显著降低。此外,BPAG1e缺陷的角质形成细胞表现出前后极性丧失,并显示出异常的运动性。通过诱导组成型活性Rac1或活性丝切蛋白的表达,这些缺陷得以挽救。这些数据表明,BPAG1e通过决定Rac1的激活,是有效调节角质形成细胞极性和迁移所必需的。